GSM6620303: NC, IL-6, biol rep1; Homo sapiens; RNA-Seq
RNAs were extracted using the RNA XS kit according to the manufacturer instruction (Macherey-Nagel, NucleoSpin RNA XS, 740902) the libraries was prepared from 10 ng of total RNA. The mRNA poly(A) tail was tagged with universal adapters, well-specific barcodes and unique molecular identifiers (UMIs) during template-switching reverse transcriptase. Barcoded cDNAs from multiple samples were then pooled, amplified and tagmented using a transposon-fragmentation approach, which enriches for 3'ends of cDNA. 38.5 nM of full-length cDNAs was used as input to the Nextera DNA Flex library prep kit (ref 20018704, Illumina), which enriches for 3′ends of cDNA. Size library is controlled on 2200 Tape Station Sytem (Agilent Technologies). A library of 350-800bp (major pic at 484 pb) was run on an Illumina NovaSeq 6000 using a NovaSeq 6000 SP Reagent Kit 100 cycles according to manufacturer's protocol (NovaSeq 6000 Sequencing System Guide Document #1000000019358v11, Illumina). 3'seq-RNA 